Experimental investigations were carried out for Cupriavidus necator (MTCC 1472)-based improved production of poly-3 hydroxy butyrate (PHB) through induced nitrogen limiting fed-batch cultivation strategies. Initially Plackett-Burman design and response surface methodology were implemented to optimize most influencing process parameters. With optimized process parameter values, continuous feeding strategies ware applied in a 5-l fermenter with table sugar concentration of 100 g/l, nitrogen concentration of 0.12 g/l for fed-batch fermentation with varying dilution rates of 0.02 and 0.046 1/h. To get enriched production of PHB, concentration of the sugar was further increased to 150 and 200 g/l in feeding. Maximum concentrations of PHB achieved were 22.35 and 23.07 g/l at those dilution rates when sugar concentration maintains at 200 g/l in feeding. At maximum concentration of PHB (23.07 g/l), productivity of 0.58 g/l h was achieved with maximum PHB accumulation efficiency up to 64% of the dry weight of biomass. High purity of PHB, close to medical grade was achieved after surfactant hypochlorite extraction method, and it was further confirmed by SEM, EDX, and XRD studies.
Keywords: Alcaligenes eutrophus; Fed-batch cultivation; Nutritional stressed condition; Poly-3-hydroxy butyrate; Response surface methodology.